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Bacterial biofilms have been demonstrated to be closely related to clinical infections and contribute to drug resistance. Berberine, which is the main component of Coptis chinensis, has been reported to have efficient antibacterial activity. This study aimed to investigate the potential effect of a combination of berberine with ciprofloxacin (CIP) to inhibit Salmonella biofilm formation and its effect on expressions of related genes (rpoE, luxS, and ompR). The fractional inhibitory concentration (FIC) index of the combination of berberine with CIP is 0.75 showing a synergistic antibacterial effect. The biofilm's adhesion rate and growth curve showed that the multi-resistant Salmonella strain had the potential to form a biofilm relative to that of strain CVCC528, and the antibiofilm effects were in a dose-dependent manner. Biofilm microstructures were rarely observed at 1/2 × MIC/FIC concentrations (MIC, minimal inhibition concentration), and the combination had a stronger antibiofilm effect than each of the antimicrobial agents used alone at 1/4 × FIC concentration. LuxS, rpoE, and ompR mRNA expressions were significantly repressed (p < 0.01) at 1/2 × MIC/FIC concentrations, and the berberine and CIP combination repressed mRNA expressions more strongly at the 1/4 × FIC concentration. The results indicate that the combination of berberine and CIP has a synergistic effect and is effective in inhibiting Salmonella biofilm formation via repression of luxS, rpoE, and ompR mRNA expressions.
Subject(s)
Anti-Infective Agents , Berberine , Biofilms , Ciprofloxacin , Coptis , Drug Combinations , Drug Resistance , Drug Resistance, Multiple , Repression, Psychology , RNA, Messenger , SalmonellaABSTRACT
Objective:To explore the clinical effect of different surgical methods combined with Neoadjuvant chemotherapy in treating patients with breast cancer.Methods:80 patients treated and diagnosed in our hospital from January,2012 to January,2014 were enrolled in the present study.According to the willingness,physical condition and financial situation,they were divided them into group A (23 cases) and group B (57 cases).Neoadjuvant chemotherapy of EC regimens (epirubicin+cyclophosphamide) was applied to both groups,on the basis of which,group A received breast-conserving surgery,group B received modified radical mastectomy.The clinical effect,breast cosmetic result,life quality,psychological states were compared in both groups.Results:The operation time,extubation time were significantly shorter than those of group B (p ＜0.05),the amount of bleeding,AMA,HRSD score,occurrence rate of complications in group A were significantly lower than those of group B (p＜0.05),the breast cosmetic result of group A was obviously better than that of group B (p＜0.05),and the QLQ-BR23 score in group A was significantly higher than that of group B (p＜0.05).The 2-year survival rate and 2 year recurrence rate showed no statistical difference between the two groups (p ＞0.05).Conclusion:Breast-conserving surgery combined with epirubicin neoadjuvant chemotherapy was effective in treating breast cancer,which could decrease the complications,improve the breast cosmetic result and quality of life.
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Objective The aim of this study was to investigate the polymorphism of SLA class II genes in Canadian SPF Yorkshire and Landrace pigs.Methods Blood samples were obtained from 15 SPF Yorkshire and 22 Landrace pigs for isolation of peripheral blood mononuclear cells respectively, and the DQB1, DRB1 and DQA genes were amplified by PCR after reverse transcription.SLA class II genes were obtained by analyzing the direct and cloning result.The polymorphism of alleles was analyzed using the DNAsp 5.0 software.Results A total of 25 alleles were identified at three genes, including eight DQB1, ten DRB1 and seven DQA, and three alleles were submitted the complete sequences for the first time.The official allele names were assigned as SLA-DQB1*0212 (KU754590), SLA-DQB1*0203 (KU754591) and DRB1*06:07(KU754601) by the SLA Nomenclature Committee.Three novel DQA alleles were discovered.Five of the 15 amino acids, one of the 16 amino acids and 11 of the 19 amino acids, which bind processing antigens, showed well conserved among the alleles of DQB1, DRB1 and DQA genes in the SPF Yorkshire and Landrace pigs, respectively.Neighbor-joining tree showed that the three genes were divided into two clusters, respectively.There was a close relationship between SPF Yorkshire and Landrace pigs and foreign Yucatan miniature pigs, and it showed no obvious genetic distance with other pigs.Conclusions A total of 25 SLA class II alleles have been identified successfully in this study, and there are more abundant polymorphism for them.There is a widely distribution for SLA class II alleles identified in this study in other pig breeds.It is critical for the eventual future use of SPF Yorkshire and Landrace pigs as classical laboratory animal models.
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Objective To determine the reproductive physiology and blood physiological and biochemical characteristics of SPF Yorkshire and Landrace swine.Methods Ten reproductive physiology parameters,19 blood physiological parameters and 18 blood biochemical parameters in SPF Yorkshire and Landrace swine were measured using conventional methods and the differences between population,between age groups and between both sexes were analyzed.Results There were no significant differences(P>0.05) in reproductive physiology parameters and most blood physiological and biochemical parameters of the SPF Yorkshire and Landrace swine.A few of parameters,such as blood physiological indices GRAN,HGB,RDW,PLT,PCT,and blood biochemical indices ALKP,CHOL,TBIL,BUN,showed significant difference(P<0.05) between populations,between age groups and between both sexes,however,the values of difference were rather small,deviated from the normal range.Conclusion The physiological and biochemical characteristics of SPF Yorkshire and Landrace swine are basically stable and there is no significant difference compared with other laboratory miniature pigs.This study will provide valuable basic data for raising velvet yield,establishment of animal models and evaluating the genetic quality of closed colony.
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In order to improve the surface properties of PLGA polymer for a better material/cell interface to modulate the cells behaviors, we prepared a novel three-block copolymer, PLGA-[ASP-PEG], and immobilized an RGD-containing peptide, Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC) on the surface of it. Transforming growth factor-beta1 (TGF-beta1) was transfected into bone marrow stromal cells (MSCs) employed as seeded cells. Cell adhesion, spreading, proliferation and differentiation on this material were investigated. The results showed that the cell adhesive ratio on RGD-modified materials was higher than on un-modified materials (P<0.05). The extent of cell spreading was also wider on RGD-modified materials than on un-modified materials. Cell proliferation indices of transfected MSCs were increased as compared with the un-transfected MSCs (P<0.05). The ALP activities in the MSCs cultured with RGD-modified materials were higher than on un-modified materials after 14 days (P<0.05), and those in transfected MSCs were higher than in un-transfected MSCs (P<0.05). It was suggested that the combined use of RGD-modification and TGF-beta gene transfection could improve the interaction of biomaterial and cells.
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In this study, the bioactivity of a novel BMP2-derived oligopeptide P24 was investigated by using the model of rabbit femoral defect after loaded in the biodegradable poly (lactic acid / glycolic acid / asparagic acid-co-polyethylene glycol) (PLGA-[ASP-PEG]). A 1.5-cm unilateral segmental bone defect was created in the left femoral diaphysis in each of the 30 new zealand white rabbits. The defects of 18 legs filled with BMP2-derived peptide P24 combined with PLGA-[ASP-PEG] scaffold serves as the experimental group, and the defects in the rest 12 rabbits filled with (PLGA-[ASP-PEG]) without P24 as control group. The bone-repairing capability in the target region of the two group was grossly, radiologically, histopathologically and biomechanically evaluated 4, 8 and 12 weeks after the operation. Our results showed that in each group, primary healing of incision was achieved in the two groups. Radiographically, in experimental group, defects were filled with induced callus within 8 weeks, and a cortical bone-like structure was observed in some animals at the 12th week. According to the standardized stage of bone defect repair, 9 (64.28%) achieved grade-4 healing. In contrast, little bone formation was seen in the defects even 12 weeks after the operation, and 5 (62.50%) had grade 0 healing in this group. Histologically, tissue engineering material was mostly absorbed and cartilage was found around implants in the experimental group at the 4th week; 8 weeks after operation, the engineering material was completely absorbed, and formation of woven bone was observed and typical trabecular bone structure could be seen. In control group, 8 weeks after operation, the defect was filled with fibrous tissues, and no bone-like structure was observed. Statistical analysis showed very significant difference in biomechanical indicators between the two groups (P<0.05). It is concluded that new oligopeptide P24 can induce excellent bone regeneration and promote bone repair.
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A 23 amino acid, bifunctional integrin-targeted synthetic oligopeptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells (BMSCs). Synthesis of the peptide (K)16GRGDSPC was performed on a solid-phase batch peptide synthesizer. BMSCs were transfected with plasmid DNA coding for luciferase by (K)16GRGDSPC and the transfection efficiency was assayed. The influences of chloroquine and polyethyleneimine on the transfection efficiency were also examined. The target specificity of (K)16GRGDSPC to mediate exogenous gene into BMSCs was analyzed using cell attachment test and gene delivery inhibition test. The results showed that the transfection efficiency of the oligopeptide vector was lower than that of Lipofectamine. But in the presence of endosomal buffer chloroquine or endosomal disrupting agent polyethyleneimine, the transfection efficiency of the vector was greatly enhanced. In addition, RGD-containing peptides inhibited BMSCs' attachment to the 96-well plates pretreated with fibronectin or vitronectin and significantly decreased the transfection efficiency of the oligopeptide vector. These studies demonstrated that oligopeptide (K)16GRGDSPC was an ideal novel targeted non-viral gene delivery vector, which was easy to be synthesized, high efficient and low cytotoxicity. The vector could effectively deliver exogenous gene into rat BMSCs.
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A 23 amino acid, bifunctional integrin-targeted synthetic oligopeptide was evaluated for ex vivo gene delivery to rabbit bone marrow stromal cells (BMSCs). Synthesis of the peptide (K)16GRGDSPC was performed on a solid-phase batch peptide synthesizer. BMSCs were transfected with plasmid DNA coding for luciferase by (K)16GRGDSPC and the transfection efficiency was assayed. The influences of chloroquine and polyethyleneimine on the transfection efficiency were also examined. The target specificity of (K)16GRGDSPC to mediate exogenous gene into BMSCs was analyzed using cell attachment test and gene delivery inhibition test. The results showed that the transfection efficiency of the oligopeptide vector was lower than that of Lipofectamine. But in the presence of endosomal buffer chloroquine or endosomal disrupting agent polyethyleneimine, the transfection efficiency of the vector was greatly enhanced. In addition, RGD-containing peptides inhibited BMSCs' attachment to the 96-well plates pretreated with fibronectin or vitronectin and significantly decreased the transfection efficiency of the oligopeptide vector. These studies demonstrated that oligopeptide (K)16GRGDSPC was an ideal novel targeted non-viral gene delivery vector, which was easy to be synthesized, high efficient and low cytotoxicity. The vector could effectively deliver exogenous gene into rat BMSCs.
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Objective To investigate the feasibility and efficiency of K16GRGDSPC(K16-RGD) for exogenous gene transfer to bone marrow derived stroma cells(BMSCs).Methods The peptide K16-RGD was synthesized by solid-phase batch peptide synthesizer.The K16-RGD was used as vector for transfecting the luciferase into BMSCs and the expression of the luciferase gene was monitored.The influence of chloroquine and polyethyleneimine was observed.The targeted specificity was examined by cell attachment test and transfection inhibitation test.Results The transfection efficiency of K16-RGD vector was lower than that of commercial lipofectamine.But the efficency of K16-RGD was greatly enhanced in the presence of chloroquine and polyethyleneimine.The peptides containing RGD inhibited the BMSCs attachment to the 96-well plates and decreased the transfection efficiency of K16-RGD significantly.Conclusion Peptide K16GRGDSPC is a new kind of targeted-nonviral gene delivery vector,which is easy to be synthesized,high efficiency and low cytotoxicity.